Table 1.
The plasmids and corresponding clones used in this study. The copy number of reporter gene in strains were obtained by means of qPCR and are represented by the mean ± SD of three independent experiments.
| Plasmids | Corresponding clones | Reporter gene copies | Mutation sites in AOX1 promoter |
|---|---|---|---|
| pPICZA | — | — | |
| pGAPZαA | — | — | |
| pPICZA-pGH | — | — | |
| pPICZA-pGH-Xba | — | — | |
| pPICZA-LacZ | — | — | |
| GAP-pPICZA-pGH | WT | 1.35 ± 0.19 | |
| GAP-pPICZA-Del-S1-pGH | Del-S1 | 1.55 ± 0.21 | Deletion of S1 tract within PAOX1 |
| GAP-pPICZA-Del-S2-pGH | Del-S2 | 0.89 ± 0.31 | Deletion of S2 tract within PAOX1 |
| GAP-pPICZA-Del-S3-pGH | Del-S3 | 1.97 ± 0.62 | Deletion of S3 tract within PAOX1 |
| GAP-pPICZA-Del-S4-pGH | Del-S4 | 1.21 ± 0.6 | Deletion of S4 tract within PAOX1 |
| GAP-pPICZA-Del-S5-pGH | Del-S5 | 1.03 ± 0.06 | Deletion of S5 tract within PAOX1 |
| GAP-pPICZA-Del-S6-pGH | Del-S6 | 1.45 ± 0.36 | Deletion of S6 tract within PAOX1 |
| GAP-pPICZA-Del-S7-pGH | Del-S7 | 0.71 ± 0.18 | Deletion of S7 tract within PAOX1 |
| GAP-pPICZA-Del-S8-pGH | Del-S8 | 0.92 ± 0.25 | Deletion of S8 tract within PAOX1 |
| GAP-pPICZA-Add-S1-pGH | Add-S1 | 1.02 ± 0.28 | Addition of 15 bp poly dA:dT tracts in S1 site |
| GAP-pPICZA-Add-S2-pGH | Add-S2 | 1.18 ± 0.21 | Addition of 15 bp poly dA:dT tracts in S2 site |
| GAP-pPICZA-Add-S3-pGH | Add-S3 | 1.02 ± 0.08 | Addition of 15 bp poly dA:dT tracts in S3 site |
| GAP-pPICZA-Add-S4-pGH | Add-S4 | 1.21 ± 0.11 | Addition of 15 bp poly dA:dT tracts in S4 site |
| GAP-pPICZA-Add-S5-pGH | Add-S5 | 1.47 ± 0.16 | Addition of 15 bp poly dA:dT tracts in S5 site |
| GAP-pPICZA-Add-S6-pGH | Add-S6 | 1.14 ± 0.06 | Addition of 15 bp poly dA:dT tracts in S6 site |
| GAP-pPICZA-Add-S7-pGH | Add-S7 | 1.05 ± 0.02 | Addition of 15 bp poly dA:dT tracts in S7 site |
| GAP-pPICZA-Add-S8-pGH | Add-S8 | 1.02 ± 0.24 | Addition of 15 bp poly dA:dT tracts in S8 site |
| GAP-pPICZA-Double-Del-S12-pGH | Double-Del-S12 | 1.04 ± 0.16 | Deletion of S1 and S2 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S24-pGH | Double-Del-S24 | 0.97 ± 0.20 | Deletion of S2 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S34-pGH | Double-Del-S34 | 1.29 ± 0.30 | Deletion of S3and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S46-pGH | Double-Del-S46 | 1.05 ± 0.31 | Deletion of S4and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S56-pGH | Double-Del-S56 | 0.86 ± 0.21 | Deletion of S5 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S57-pGH | Double-Del-S57 | 1.38 ± 0.06 | Deletion of S5 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S58-pGH | Double-Del-S58 | 0.83 ± 0.40 | Deletion of S5 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S67-pGH | Double-Del-S67 | 1.27 ± 0.47 | Deletion of S6 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S78-pGH | Double-Del-S78 | 0.80 ± 0.03 | Deletion of S7 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S12-pGH | Double-Add-S12 | 1.27 ± 0.23 | Addition of 15 bp poly dA:dT tract in S1 and S2 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S24-pGH | Double-Add-S24 | 1.27 ± 0.12 | Addition of 15 bp poly dA:dT tract in S2 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S34-pGH | Double-Add-S34 | 0.84 ± 0.09 | Addition of 15 bp poly dA:dT tract in S3 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S46-pGH | Double-Add-S46 | 1.01 ± 0.17 | Addition of 15 bp poly dA:dT tract in S4 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S56-pGH | Double-Add-S56 | 0.92 ± 0.19 | Addition of 15 bp poly dA:dT tract in S5 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S57-pGH | Double-Add-S57 | 1.15 ± 0.30 | Addition of 15 bp poly dA:dT tract in S5 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S58-pGH | Double-Add-S58 | 1.28 ± 0.27 | Addition of 15 bp poly dA:dT tract in S5 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S67-pGH | Double-Add-S67 | 1.13 ± 0.12 | Addition of 15 bp poly dA:dT tract in S6 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S78-pGH | Double-Add-S78 | 1.38 ± 0.45 | Addition of 15 bp poly dA:dT tract in S7 and S8 tracts within PAOX1 |
| GAP-pPICZA-LacZ | WT-L | 1.17 ± 0.05 | |
| GAP-pPICZA-Del-S1-LacZ | Del-S1-L | 1.04 ± 0.25 | Deletion of S1 tract within PAOX1 |
| GAP-pPICZA-Del-S2-LacZ | Del-S2-L | 1.19 ± 0.3 | Deletion of S2 tract within PAOX1 |
| GAP-pPICZA-Del-S3-LacZ | Del-S3-L | 2.07 ± 0.58 | Deletion of S3 tract within PAOX1 |
| GAP-pPICZA-Del-S4-LacZ | Del-S4-L | 1.19 ± 0.20 | Deletion of S4 tract within PAOX1 |
| GAP-pPICZA-Del-S5-LacZ | Del-S5-L | 1.37 ± 0.21 | Deletion of S5 tract within PAOX1 |
| GAP-pPICZA-Del-S6-LacZ | Del-S6-L | 1.11 ± 0.23 | Deletion of S6 tract within PAOX1 |
| GAP-pPICZA-Del-S7-LacZ | Del-S7-L | 1.12 ± 0.02 | Deletion of S7 tract within PAOX1 |
| GAP-pPICZA-Del-S8-LacZ | Del-S8-L | 0.86 ± 0.23 | Deletion of S8 tract within PAOX1 |
| GAP-pPICZA-Add-S1-LacZ | Add-S1-L | 0.77 ± 0.01 | Addition of 15 bp poly dA:dT tracts in S1 site |
| GAP-pPICZA-Add-S2-LacZ | Add-S2-L | 0.79 ± 0.30 | Addition of 15 bp poly dA:dT tracts in S2 site |
| GAP-pPICZA-Add-S3-LacZ | Add-S3-L | 1.02 ± 0.10 | Addition of 15 bp poly dA:dT tracts in S3 site |
| GAP-pPICZA-Add-S4-LacZ | Add-S4-L | 1.14 ± 0.31 | Addition of 15 bp poly dA:dT tracts in S4 site |
| GAP-pPICZA-Add-S5-LacZ | Add-S5-L | 0.86 ± 0.01 | Addition of 15 bp poly dA:dT tracts in S5 site |
| GAP-pPICZA-Add-S6-LacZ | Add-S6-L | 1.73 ± 0.28 | Addition of 15 bp poly dA:dT tracts in S6 site |
| GAP-pPICZA-Add-S7-LacZ | Add-S7-L | 0.88 ± 0.01 | Addition of 15 bp poly dA:dT tracts in S7 site |
| GAP-pPICZA-Add-S8-LacZ | Add-S8-L | 0.88 ± 0.21 | Addition of 15 bp poly dA:dT tracts in S8 site |
| GAP-pPICZA-Double-Del-S12-LacZ | Double-Del-S12-L | 1.15 ± 0.26 | Deletion of S1 and S2 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S24-LacZ | Double-Del-S24-L | 1.50 ± 0.02 | Deletion of S2 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S34-LacZ | Double-Del-S34-L | 1.30 ± 0.37 | Deletion of S3and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S46-LacZ | Double-Del-S46-L | 1.10 ± 0.16 | Deletion of S4and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S56-LacZ | Double-Del-S56-L | 1.06 ± 0.19 | Deletion of S5 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S57-LacZ | Double-Del-S57-L | 1.27 ± 0.12 | Deletion of S5 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S58-LacZ | Double-Del-S58-L | 0.98 ± 0.06 | Deletion of S5 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S67-LacZ | Double-Del-S67-L | 1.19 ± 0.25 | Deletion of S6 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Del-S78-LacZ | Double-Del-S78-L | 1.50 ± 0.34 | Deletion of S7 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S12-LacZ | Double-Add-S12-L | 1.05 ± 0.01 | Addition of 15 bp poly dA:dT tract in S1 and S2 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S24-LacZ | Double-Add-S24-L | 1.01 ± 0.28 | Addition of 15 bp poly dA:dT tract in S2 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S34-LacZ | Double-Add-S34-L | 0.90 ± 0.27 | Addition of 15 bp poly dA:dT tract in S3 and S4 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S46-LacZ | Double-Add-S46-L | 0.87 ± 0.19 | Addition of 15 bp poly dA:dT tract in S4 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S56-LacZ | Double-Add-S56-L | 0.94 ± 0.23 | Addition of 15 bp poly dA:dT tract in S5 and S6 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S57-LacZ | Double-Add-S57-L | 1.29 ± 0.15 | Addition of 15 bp poly dA:dT tract in S5 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S58-LacZ | Double-Add-S58-L | 0.83 ± 0.07 | Addition of 15 bp poly dA:dT tract in S5 and S8 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S67-LacZ | Double-Add-S67-L | 0.86 ± 0.17 | Addition of 15 bp poly dA:dT tract in S6 and S7 tracts within PAOX1 |
| GAP-pPICZA-Double-Add-S78-LacZ | Double-Add-S78-L | 0.86 ± 0.20 | Addition of 15 bp poly dA:dT tract in S7 and S8 tracts within PAOX1 |